Team:Stuttgart/Contribution

Contributions Contributions

Contributions

Our project focused on the damage suppressor protein (dsup) from tardigrades. Our goal was to achieve the expression of the protein in E. coli and HeLa cells to make them more resistant to UV radiation. In addition, we also wanted to purify the dsup to investigate if it also protects the cells when it is only present in their environment. To achieve the expression and subsequent purification of the previously poorly studied dsup in the cells we had to design new plasmids.

For our experiments we designed a total of 4 plasmids. Three of them are for E. coli, because we need a His-tag for the purification, but we don't know how this affects the function of the protein. So we designed a plasmid with His-tag at the C-terminus, a plasmid with His-tag at the N-terminus and a plasmid without a His-tag. The last plasmid was designed for HeLa cells, it has an N-terminal His-tag. The exact designs can be found in the Wet-Lab section.

Since unfortunately only competent E. coli cells could be generated, the effect of dsup on intracellular resistance to UV radiation was only investigated in these cells. In HeLa cells, the change in resistance to extracellular dsup was then investigated using a live cell imaging experiment. The resulting findings can be read in the results section.

We hope our newly designed plasmids and the results from our irradiation experiments will help future teams to further investigate the damage suppressor protein from tardigrades.